Journal: Aging Cell

Article Title: The Proinflammatory Secretome of Senescent Cells Can Be Controlled by a HIF2A ‐Dependent Upregulation and a FURIN ‐Dependent Cleavage of the ANGPTL4 Secreted Factor

doi: 10.1111/acel.70307

Figure Lengend Snippet: Cleavage of ANGPTL4 by FURIN is required for its ability to promote the proinflammatory SASP. (A‐C) MRC5/RAF:ER cells were transfected with control (siCTRL) or FURIN (siFURIN) siRNA and treated (+) or not (−) with 4‐OHT to induce senescence (OIS). (A) RT‐qPCR analysis against the indicated genes 3 days after 4‐OHT treatment. Mean ± SEM of n = 3 independent experiments. Paired one‐way ANOVA test results are shown. (B) Western blot analysis of FURIN, ANGPTL4: Full length in whole cell extracts (ANGPTL4) and secreted in the supernatant (cANGPTL4) and IL6. Representative picture of n = 3 independent experiments. (C) RT‐qPCR analysis of the indicated SASP encoding genes. Mean ± SEM of n = 3 independent experiments. Paired one‐way ANOVA test values are shown. (D) Relative mRNA expression of FURIN , ANGPTL4 and proinflammatory SASP encoding genes measured by RT‐qPCR in MRC5 cells overexpressing or not ANGPTL4 (ANGPTL4 OE) and transfected with control (siCTRL) or FURIN (siFURIN) siRNA. Mean ± SEM of n = 4 independent experiments. Paired one‐way ANOVA test results are shown.

Article Snippet: The membranes were then blocked for 1 h with 5% milk in TBS‐T 0.05% and subsequently incubated overnight at 4°C with primary antibodies in TBS‐T with 5% milk: α‐ANGPTL4 (sc‐373761, Santa Cruz, 1/250), α‐IL6 (sc‐28343, Santa Cruz, 1/500), α‐FURIN (18413‐1‐AP, Proteintech, 1/1000), α‐TUBULIN (T6199‐100, Sigma, 1/5000). α‐HIF1A (610958, BD Biosciences, 1/1000), HIF2A (NB100‐132, Novus Biologicals, 1/1000), INHBA (GTX108405, GeneTex 1/1000).

Techniques: Transfection, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Renal Failure

Article Title: Urotensin II system contributes to ischemic acute kidney injury in neonatal pigs

doi: 10.1080/0886022X.2025.2534018

Figure Lengend Snippet: Upregulation of the UII system in the kidneys of neonatal pigs following IR injury. Quantitative RT-PCR analysis showing relative mRNA expression levels of (A) furin, (B) UII, (C) URP, and (D) UT in whole kidney tissue, and (E) UT in intrarenal arteries from sham and IR piglets ( n = 4 per group). (F–G) Representative Western blot and densitometric analysis of UT protein expression in intrarenal arteries ( n = 4 per group). (H–I) Western blot and quantification of furin protein expression in kidney tissues ( n = 4 per group). (J–K) representative immunohistochemistry (IHC) images and quantification of UII immunostaining intensity in renal tubules from sham and IR piglets (4 kidney sections per group). * p < 0.05, unpaired t -test.

Article Snippet: For Western blot analysis, the primary antibodies included a rabbit polyclonal anti-furin antibody (18413-1-AP, Proteintech, Rosemont, IL, USA) at a 1:500 dilution and a rabbit polyclonal anti-GPR14 (UT receptor) antibody (TA358466, OriGene Technologies, Rockville, MD, USA) at a 1:100 dilution.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemistry, Immunostaining

Journal: Renal Failure

Article Title: Urotensin II system contributes to ischemic acute kidney injury in neonatal pigs

doi: 10.1080/0886022X.2025.2534018

Figure Lengend Snippet: Furin inhibition reduces cIR-driven UII production, and UT and furin blockade attenuate cIR-induced cytotoxicity in primary neonatal pig proximal tubule epithelial cells. (A) Representative fluorescence microscopy images showing furin and UII immunostaining and their colocalization in primary neonatal pig proximal tubule epithelial cells (PTECs; 3 biological replicates). (B) Quantification of intracellular furin levels in control (ctrl) and chemical ischemia-reperfusion (cIR)–treated cells ( n = 4). (C) Secreted UII levels in cIR-treated cells in the absence or presence of the furin inhibitor SSM3 trifluoroacetate (SSM3; n = 4). (D) cIR-induced cytotoxicity was significantly reduced by pharmacological inhibition of furin (SSM3) and by UT inhibition with urantide (URTD). * p < 0.05 ctrl vs. cIR; p < 0.05 cIR vs. SSM3 + cIR; $ p < 0.05 cIR vs. both SSM3 + cIR and URTD + cIR. Unpaired t -test (B); One-way ANOVA and Holm-Šídák’s multiple comparisons tests (C–D); n = 4, each. Scale bar = 50 µm.

Article Snippet: For Western blot analysis, the primary antibodies included a rabbit polyclonal anti-furin antibody (18413-1-AP, Proteintech, Rosemont, IL, USA) at a 1:500 dilution and a rabbit polyclonal anti-GPR14 (UT receptor) antibody (TA358466, OriGene Technologies, Rockville, MD, USA) at a 1:100 dilution.

Techniques: Inhibition, Fluorescence, Microscopy, Immunostaining, Control